Photosynth Res (2010) 106:33–46
DOI 10.1007/s11120-010-9538-8
REVIEW
State transitions at the crossroad of thylakoid signalling pathways
Sylvain Lemeille • Jean-David Rochaix
Received: 21 December 2009 / Accepted: 11 February 2010 / Published online: 9 March 2010
Ó Springer Science+Business Media B.V. 2010
Abstract In order to maintain optimal photosynthetic
activity under a changing light environment, plants and
algae need to balance the absorbed light excitation energy
between photosystem I and photosystem II through pro-
cesses called state transitions. Variable light conditions
lead to changes in the redox state of the plastoquinone pool
which are sensed by a protein kinase closely associated
with the cytochrome b6f complex. Preferential excitation of
photosystem II leads to the activation of the kinase which
phosphorylates the light-harvesting system (LHCII), a
process which is subsequently followed by the release of
LHCII from photosystem II and its migration to photo-
system I. The process is reversible as dephosphorylation of
LHCII on preferential excitation of photosystem I is fol-
lowed by the return of LHCII to photosystem II. State
transitions involve a considerable remodelling of the thy-
lakoid membranes, and in the case of Chlamydomonas,
they allow the cells to switch between linear and cyclic
electron flow. In this alga, a major function of state tran-
sitions is to adjust the ATP level to cellular demands.
Recent studies have identified the thylakoid protein kinase
Stt7/STN7 as a key component of the signalling pathways
of state transitions and long-term acclimation of the pho-
tosynthetic apparatus. In this article, we present a review
on recent developments in the area of state transitions.
Electronic supplementary material The online version of this
article (doi:10.1007/s11120-010-9538-8) contains supplementary
material, which is available to authorized users.
S. Lemeille  J.-D. Rochaix (&)
Departments of Molecular Biology and Plant Biology,
University of Geneva, 30, Quai Ernest Ansermet,
1211 Geneva, Switzerland
e-mail: Jean-David.Rochaix@unige.ch
Keywords Photosynthesis  State transitions 
LHCII kinase  Arabidopsis  Chlamydomonas
Introduction
The primary reactions of photosynthesis occur in the thy-
lakoid membranes and are catalyzed by the photosystem II
(PSII) and photosystem I (PSI) complexes, and their
associated antennae. These complexes act in series and are
linked together through the plastoquinone pool, the cyto-
chrome b6f complex and plastocyanin within the photo-
synthetic electron transport chain. The collected light
energy is channelled from the antennae to the reaction
centres of PSII and PSI and used to induce stable charge
separations across the thylakoid membrane with sub-
sequent oxidation of water by PSII, followed by multiple
electron transfer reactions, and photoreduction of ferre-
doxin by PSI. Ultimately, this process creates reducing
power and a proton gradient across the membrane which is
used by ATP synthase to produce ATP. The differentiation
of thylakoids into grana and stroma lamellar regions is
believed to reflect the uneven distribution of the photo-
synthetic complexes. Whereas PSI and the ATP synthase
with their bulky stromal domains are localized in the non-
appressed regions consisting of the stroma lamellae and the
grana end membranes and margins, PSII and its light-
harvesting system (LHCII) are embedded in the appressed
regions of the grana.
The photosynthetic machinery is remarkably dynamic
and has developed several mechanisms to adapt to both
high- and low-light conditions. In nature, photosynthetic
organisms are constantly subjected to changes in their light
environment, and they need to adjust their photosynthetic
activity accordingly. Under high light when the absorbed
123
34
light energy exceeds the capacity of the photosynthetic
apparatus, the excess absorbed energy is dissipated through
heat, a process called non-photochemical quenching (for
review, see Holt et al. 2004). At the same time, the pho-
tosynthetic system is susceptible to light-induced damage
leading to photoinhibition and repair at the level of PSII.
Because the antenna systems of PSII and PSI have a
different composition and hence different light absorption
properties, their excitation can be unbalanced under
changing light conditions. Thus, when PSII is preferentially
excited by light absorbed mostly by PSII, the plastoquinone
(PQ) pool is reduced, and on docking of plastoquinol
(PQH2) to the Qo site of the cytochrome b6f complex, a
protein kinase is activated that phosphorylates LHCII
(Fig. 1A). This event triggers the release of LHCII from
PSII and its migration to PSI thereby rebalancing the light
excitation energy between the two photosystems (state 2).
The process is reversible as preferential excitation of PSI
by light enriched in the far-red region induces the
dephosphorylation of LHCII and its return to PSII (state 1).
This rebalancing processes called state transitions also
involve a remodelling of the thylakoid membranes and lead
to an optimal utilization of the absorbed light energy.
State transitions have been studied in several photo-
synthetic organisms including algae, land plants and cya-
nobacteria. In this article, we review recent studies on state
transitions performed mainly in the unicellular green alga
Chlamydomonas reinhardtii and the model plant Arabid-
opsis thaliana. These two organisms have emerged as very
powerful and complementary model systems in which
genetic, biochemical, physiological and biophysical
approaches can be coupled efficiently. It is of particular
interest to compare the acclimation of a mobile unicellular
alga with that of a sessile multicellular plant, and this also
provides interesting insights into evolutionary aspects of
light acclimation. Earlier studies on state transitions have
been extensively discussed in several reviews on state
transitions (Allen 1992; Wollman 2001; Rochaix 2007).
STN7/Stt7 protein kinase is the key player for state
transitions
State transitions were discovered independently by Murata
(1969) and Bonaventura and Myers (1969) while they were
studying the fluorescence properties of two unicellular
organisms, the red alga Porphyridium cruentum in one
case, and the green alga Chlorella pyrenoidosa, in the
other. These investigators subjected these algae to light
absorbed preferentially by PSII and found that, within a
few minutes, PSI absorbed more light excitation energy. In
contrast, illumination with light absorbed preferentially by
PSI was followed by an increased light absorption by PSII.
123
Photosynth Res (2010) 106:33–46
The increase in the light absorption capacity of PSI and
PSII following these changes in light conditions were
referred to as state 2 and state 1, respectively. It was later
found that regulation of excitation energy transfer from
PSII to PSI occurred through phosphorylation of the light-
harvesting complex of PSII (LHCII) and that this process
was associated with fluorescence changes (Bennett 1977,
1979). The kinase involved in this phosphorylation was
shown to be activated through reduction of the plastoqui-
none pool after preferential excitation of PSII (Allen et al.
1981; Horton and Black 1980). The finding, that the two
photosystems are not equally distributed in the thylakoid
membranes with PSII and PSI localized mainly in the
grana and stromal lamellae, respectively (Andersson and
Andersson 1980), implied that the light excitation energy
needs to be redistributed between these thylakoid regions
and involves the migration of LHCII between PSII and PSI
(Delosme et al. 1996).
A key finding was that the cytochrome b6f complex
plays an important role in state transitions. Mutants of
C. reinhardtii lacking this complex are unable to phos-
phorylate LHCII and are locked in state 1 (Wollman and
Lemaire 1988). Further analysis revealed that it is not the
redox state of the plastoquinone pool per se which is critical
for the activation of the kinase, but the docking of plas-
toquinol to the Qo site on the luminal side of the cytochrome
b6f complex (Vener et al. 1997; Zito et al. 1999).
In spite of the fact that it was possible to identify several
kinase activities associated with thylakoids, attempts to
isolate the LHCII protein kinase by biochemical means
failed (Coughlan and Hind 1986, 1987; Sokolenko et al.
1995). Taking advantage of the fact that the N-terminal
region of LHCII contains the residues that are phosphor-
ylated during a state 1 to state 2 transition, Kohorn and
colleagues used a genetic screen for proteins that interact
with this region (Snyders and Kohorn 2001). This screen
led to the discovery of the TAK protein kinases which form
a small family related to the human TGFb1 receptor
(Snyders and Kohorn 1999). The TAK kinase was shown to
be associated with PSII and the cytochrome b6f complex.
Moreover, a decline in kinase level in antisense TAK lines
led to a decrease in LHCII phosphorylation and to a partial
deficiency in state transitions, and to light sensitivity
(Snyders and Kohorn 2001). The fact that no TAK ortholog
could be identified in Chlamydomonas suggests that the
TAK kinases perform a role which is specific to land
plants. Alternatively, these kinases may have diverged
considerably between algae and plants. Surprisingly, no
further studies have been performed on the TAK kinases
since the early studies of Kohorn.
Chlamydomonas reinhardtii has proven to be especially
attractive for a genetic approach for studying state transi-
tions because transition from state 1 to state 2 is associated
Photosynth Res (2010) 106:33–46
Fig. 1 Model for state transitions in the thylakoid membrane. A The
redox state of the PQ pool determines state transitions. When PSI is
preferentially excited, the PQ pool is oxidized. In this state, called
state 1, LHCIIs are bound to PSII. Monomeric LHCIIs (CP29, CP26
and Lhcbm5) act as connectors between PSII core and the trimeric
LHCII, and the photosynthetic electron flow proceeds in a linear
mode generating both a proton gradient across the thylakoid
membrane which is used for ATP production, and NADPH. In
contrast, when PSII is preferentially excited, the PQ pool is reduced.
Docking of plastoquinol to the Qo site of the cytochrome b6f leads to
the activation of the Stt7/STN7 kinase which is required for the
phosphorylation of CP29, CP26 and Lhcbm5 and the LHCII trimers,
and to the displacement of LHCII from PSII to PSI (state 2). The
docking of LHCII occurs on the PsaH side of PSI whereas the LHCI
belt is on the other side of PSI. In state 2 in C. reinhardtii, PSII is
35
disconnected from the electron transport chain which now operates in
a cyclic mode between PSI and the PQ pool generating mostly ATP.
State 2 can also be induced under anaerobic conditions through the
chloro-respiratory electron transport chain. B Model for inactivation
of Stt7/STN7 under high light. Reducing equivalents are shuttled
from ferredoxin and thioredoxin through a trans-thylakoid thiol-
reducing pathway mediated by CcdA and Hcf164 across the thylakoid
membrane and could reduce the disulphide bond in the lumenal
N-terminal domain of Stt7/STN7 and thereby inactivate the kinase.
P680, PSII reaction centre chlorophyll dimer; QA, QB, primary and
secondary electron acceptors of PSII; PQ/PQH2, plastoquinone/
plastoquinol, P700, PSI reaction centre chlorophyll dimer; FX, FA,
FB, 4Fe–4S centres acting as electron acceptors within PSI; PC
plastocyanin, Fd ferredoxin
123
36
with a large decrease in fluorescence in this alga (Wollman
and Delepelaire 1984). A screen based on this differential
fluorescence signal was used for isolating mutants deficient
in state transitions (Fleischmann et al. 1999; Kruse et al.
1999). In this way, the stt7 mutant was isolated, which is
blocked in state 1 and fails to phosphorylate LHCII under
state 2 conditions (Fleischmann et al. 1999). The gene
affected in stt7 is a thylakoid protein kinase called Stt7
which comprises 499 amino acids (Depège et al. 2003).
The kinase is localized in the chloroplast thylakoid mem-
branes and shown to be essential for LHCII phosphorylation
and state transitions (Depège et al. 2003). This protein dis-
plays significant sequence similarity (29% sequence iden-
tity, 43% sequence similarity) with another protein kinase
named Stl1. Both Stt7 and Stl1 have orthologs in Arabid-
opsis, called STN7 (At1g68830) and STN8 (At5g01920),
which are involved in state transitions and in PSII core
protein phosphorylation, respectively (Bellafiore et al. 2005;
Bonardi et al. 2005; Vainonen et al. 2005). Stt7/STN7-like
protein kinases are also found in other land plants, trees and
marine algae (Fig. 2, Supplementary Fig. 1).
The Stt7 kinase is required for LHCII phosphorylation
during a transition from state 1 to state 2. Characteriza-
tion of the Stt7 kinase revealed that it contains a trans-
membrane domain separating its stroma-exposed catalytic
domain from its lumen-located N-terminal end. Two
conserved cysteine residues near the N-terminus of Stt7
are critical for its activity (Lemeille et al. 2009). In
addition, coimmunoprecipitation assays showed that Stt7
interacts with LHCII proteins and cytochrome b6f com-
plex subunits, and also with PSI (Lemeille et al. 2009).
Interaction between Stt7 kinase and cytochrome b6f
complex was expected since state transitions depend
critically on this complex (Wollman 2001). Moreover,
kinase activity was detected in purified fractions of the
cytochrome b6f complex (Gal et al. 1990). Further anal-
ysis revealed that Stt7 interacts with the Rieske protein of
the cytochrome b6f complex (Lemeille et al. 2009).
Structural studies of the mitochondrial bc1 and chloro-
plast b6f complexes showed that electron transfer between
plastoquinol at the Qo site and cytochrome f is mediated
by the Rieske protein which moves from a proximal
position when the Qo site is occupied by PQH2 to a distal
position when the Qo site is unoccupied (Breyton 2000;
Stroebel et al. 2003; Zhang et al. 1998). One interesting
possibility is that this dynamic behaviour of the Rieske
protein is coupled with the activation of the Stt7 kinase
(Finazzi et al. 2001). Such a dynamic model is compatible
with the low abundance of the Stt7 kinase with a molar
ratio of 1:20 relative to the cytochrome b6f complex
(Lemeille et al. 2009). The active kinase would need to
phosphorylate several substrates before it returns to its
inactive state. The low level of Stt7 was probably the
123
Photosynth Res (2010) 106:33–46
reason of the failure of all the previous biochemical
attempts to purify the LHCII kinase.
An intriguing component of the cytochrome b6f complex
is its single chlorophyll a molecule whose chlorine ring lies
between helices F and G of the PetD subunit, whereas the
phytol chain protrudes near the Qo site (Stroebel et al.
2003). Interestingly, petD mutants affected in the binding
of chlorophyll a, besides having reduced cytochrome b6f
turnover, also display a delay in the transition from state 1
to state 2 as well as in protein phosphorylation indicating a
slower activation of the LHCII kinase by the cytochrome
b6f complex (de Lacroix de Lavalette et al. 2008). This
region of PetD may thus either form a site for interaction
with the N-terminal domain of Stt7 or it could act as a
sensor for the presence of PQH2 at the Qo site and initiate a
signalling pathway through the chlorophyll a molecule
towards the catalytic domain of Stt7 on the stromal side of
the thylakoid membrane. Additional evidence for the
importance of this region for the activation of the LHCII
kinase comes form the analysis of a Chlamydomonas strain
in which the petD and petL genes are fused to each other
resulting in a chimeric protein in which the C-terminus of
subunit IV (PetD) is fused to the N-terminus of PetL (Zito
et al. 2002). Although this strain was unimpaired in the
Q-cycle, it was unable to perform state transitions. A role
of PetL in the activation of the kinase seems unlikely
because mutants lacking PetL are still able to undergo state
transitions (Zito et al. 2002). It is thus more likely that
subunit IV is involved in this process either directly or
indirectly.
The PetO subunit of the C. reinhardtii b6f complex is
known to be phosphorylated during state transitions (Hamel
et al. 2000). In the presence of tridecyl-stigmatellin, an
inhibitor of electron transport which binds to the Qo site, and
which prevents phosphorylation of LHCII, the PetO subunit
is still phosphorylated under state 2 conditions, indicating
that it is probably the first protein to be phosphorylated upon
activation of the LHCII kinase (Finazzi et al. 2001). PetO
phosphorylation could be part of the signalling pathway of
state transitions and may act at an early step. However, no
ortholog of this protein could be found in land plants sug-
gesting that this protein is specific to Chlamydomonas.
The N-terminal region of Stt7 contains two cysteine
residues that are critical for its activity (Lemeille et al.
2009). These two cysteine residues are the only conserved
cysteine residues between Stt7 and its ortholog STN7 in
Arabidopsis thaliana (Depège et al. 2003). Moreover, these
two cysteine residues are conserved in all Stt7/STN7
orthologs (Supplementary Fig. 1). It was reported that, in
land plants, high-light treatment inactivates the LHCII
kinase through the ferredoxin–thioredoxin system (Rinta-
maki et al. 1997; Hou et al. 2003). The two conserved
cysteine could therefore be the targets of this redox system.
Photosynth Res (2010) 106:33–46 37

Fig. 2 Sequence comparison of

the Stt7 kinase from
Chlamydomonas reinhardtii and
STN7 from Arabidopsis
thaliana. The arrow indicates
the predicted cleavage site of
the chloroplast transit sequence
of Stt7. The hydrophobic region
is marked by a grey box above
the amino acid sequence. The
kinase catalytic domain is
indicated by an open box above
the amino acid sequence. The
asterisks indicate the conserved
cysteine residues that are
potential targets for the
ferredoxin–thioredoxin system.
Identified phosphorylated serine
and threonine in Stt7 and STN7
are marked by # above and
below the sequence,
respectively

# #

It thus appears likely that the activity of the STN7/Stt7
kinase is regulated by a complex network involving
cooperative redox control by PQ and the cytochrome b6f
complex as well as by the ferredoxin–thioredoxin system in
the stroma. Given the fact that these two cysteine residues
are located in the lumen whereas ferredoxin and thiore-
doxin are present in the stroma, the question arises how the
activity of the kinase is regulated under these conditions.
Recently, at least two components of a trans-thylakoid
thiol-reducing pathway have been identified in chloro-
plasts. The CcdA protein which belongs to the DsbD/DipZ
family of membrane polytopic proteins is involved in
transmembrane transfer of thiol reducing equivalents from
the stroma to the lumen (Page et al. 2004). The second

123
38
component Hcf164, a transmembrane protein containing a
thioredoxin domain in the thylakoid lumen with disulphide
reductase activity, is involved in cytochrome b6f assembly
(Lennartz et al. 2001; Motohashi and Hisabori 2006). Thus,
it is possible that the ferredoxin–thioredoxin system regu-
lates the redox state of the two Cys across the membrane
through CcdA/Hcf164 (Fig. 1B). Another possibility based
on the observation that Stt7 forms a dimer in Chlamydo-
monas (S. Lemeille and J.D. Rochaix, unpublished results)
is that these two conserved cysteine residues of Stt7 could
form two disulphide bridges between the Stt7 monomers
and that dimerization of the kinase is linked to its
activation.
Regulation of Stt7 also appears to occur at the level of
protein accumulation. Time-course experiments showed
that the level of Stt7 protein decreases under prolonged
state 1 conditions and also after high-light treatment
(Lemeille et al. 2009). Thus, Stt7 is less stable under
conditions when the kinase is in an inactive state. The
decline of the amount of Stt7 could be prevented by
addition of inhibitors of cysteine proteases. It is possible
that under state 2 conditions, Stt7 is protected from pro-
teases either because of posttranslational modifications or
by its association with other proteins. At this time, there is
no clear evidence for the presence of cysteine proteases in
the chloroplast. However, other proteases, such as Deg
proteases which are known to degrade thylakoid membrane
proteins (Huesgen et al. 2009; Adam and Clarke 2002), are
sensitive to inhibitors of cysteine proteases (Helm et al.
2007).
State transitions are associated with the phosphorylation
of LHCII (Bennett 1977), and Stt7 is essential for these
phosphorylation events (Depège et al. 2003). However, it is
still not yet known whether this kinase directly phosphor-
ylates LHCII or whether it is part of a kinase cascade
involved in the signalling pathways of state transitions.
Mapping of in vivo protein phosphorylation sites in thy-
lakoid membranes of wild-type C. reinhardtii cells under
state 1 and state 2 conditions, or when exposed to high
light, identified 19 in vivo phosphorylation sites corre-
sponding to 15 polypeptides (Turkina et al. 2006). This
study revealed that the major LHCII proteins Lhcbm4,
Lhcbm6, Lhcbm9 and Lhcbm11 are phosphorylated at Thr3
and Thr7. Because of their identical phosphopeptide
sequences, it is not possible to distinguish whether all or
only some of these proteins are phosphorylated.
Comparison of the thylakoid phosphoproteome of wild
type and stt7 mutant under state 1 and state 2 conditions
revealed that, in state 2, several Stt7-dependent phospho-
rylations of specific Thr residues occur in Lhcbm1/
Lhcbm10, Lhcbm4/Lhcbm6/Lhcbm8/Lhcbm9, Lhcbm3,
Lhcbm5, and CP29 located at the interface between PSII and
its light-harvesting system (Lemeille et al. 2010). Because
123
Photosynth Res (2010) 106:33–46
of its pivotal role in state transitions, the phosphorylation
patterns of CP29 are particularly interesting. Of the two
phosphorylation sites detected specifically in CP29 under
state 2 conditions, one is Stt7 dependent. This phosphory-
lation may play a crucial role in the dissociation of CP29
from PSII and/or in its association to PSI where it serves as a
docking site for LHCII in state 2 (Lemeille et al. 2010). Stt7
itself is phosphorylated under state 2 conditions (Lemeille
et al. 2010). However, it is not known whether this is due to
autophosphorylation or whether another kinase is involved.
State transitions and cyclic electron flow
A major role of state transitions in Chlamydomonas is to
adjust the ATP level to cellular demands. This probably
explains why the mobile LHCII represents such a large
portion of the light-harvesting system of PSII (Bulté et al.
1990). When the cellular level of ATP is low, glycolysis is
stimulated through the Pasteur effect, and the reducing
power is channelled through the chlororespiratory chain
into the plastoquinone pool, thereby activating the LHCII
kinase and promoting a transition from state 1 to state 2.
This transition triggers a switch from linear to cyclic
electron flow as indicated by the fact that in state 2 the
PSII-specific inhibitor DCMU no longer prevents the
re-reduction of the cytochrome b6f complex (Finazzi et al.
1999). In striking contrast to the wild type, under the same
state 2 conditions, DCMU blocked the re-reduction of the
cytochrome b6f complex in the stt7 mutant indicating that
this process observed in the wild type is not due to
increased influx of reducing power from the chlororespi-
ratory chain, but due to cyclic electron flow (Finazzi et al.
2002).
Such a coupling between state transitions and cyclic
electron flow is not apparent in Arabidopsis. Although it
was assumed earlier that cyclic electron flow may not be
important for steady state photosynthesis in C3 plants, this
view has changed recently (reviewed in Joliot and Joliot
2006). It now appears that the principal role of cyclic
electron flow is to match the requirements of the ATP/
NADPH ratio of 3/2 required for driving the Calvin–Ben-
son cycle especially during changes in light conditions or
under stress. Because linear electron flow gives rise to a
lower ratio, it has been estimated that there must be 20%
more PSI than PSII to allow for the recycling of one
electron in five through cyclic electron transfer (Allen
2003). The cyclic pathway transfers electrons back from
ferredoxin to the plastoquinone pool either through the
NAD(P)H dehydrogenase-dependent pathway or through
the cytochrome b6f complex using a Q-cycle-derived
mechanism (reviewed in Joliot and Joliot 2006; Shikanai
2007). Alternatively, a ferredoxin-plastoquinone reductase
Photosynth Res (2010) 106:33–46
has been proposed for direct reduction of plastoquinone by
ferredoxin (Cleland and Bendall 1992; Moss and Bendall
1984). However, this enzyme has not yet been isolated.
Two novel factors involved in cyclic electron flow, PGR5
and PGRL1, have been identified in Arabidopsis through
forward and reverse genetic screens (Munekage et al. 2002;
DalCorso et al. 2008). Plants deficient in either of these
proteins display perturbations in cyclic electron flow. Both
proteins interact physically with each other and bind to PSI
suggesting that the PGRL1–PGR5 complex may facilitate
cyclic electron flow in close association with PSI.
Role of STN7/Stt7 kinase in other processes
In response to changes in light conditions, photosynthetic
organisms induce a short-term response such as state
transitions. If preferential excitation of PSII or PSI is
maintained for longer periods, long-term acclimation
occurs which leads to changes in the amount of the
antennae proteins of PSII and PSI and to a readjustment of
photosystem stoichiometry which can be measured by the
chlorophyll a/b ratio or the fluorescence parameter FS/FM
(ratio between steady-state fluorescence and maximum
fluorescence) (Dietzel et al. 2008). This process is called
long-term response (LTR) and is achieved through a sig-
nalling network involving coordinate gene expression in
the nucleus and chloroplast (Pfannschmidt 2003). When
wild-type plants are illuminated for several days by light
preferentially absorbed by PSI relative to PSII, the exci-
tation pressure of PSII, as measured by the chlorophyll
fluorescence parameter FS/FM, increases as a result of its
increased antenna size, and the chlorophyll a/b ratio
decreases because the PSII antenna is enriched in chloro-
phyll b. The opposite occurs when the plants are illumi-
nated with PSII light. Bonardi et al. (2005) and Tikkanen
et al. (2006) observed that under all light conditions, the
response of the stn7 mutant is typical of plants acclimated
to PSI light indicating that STN7 is also required for LTR.
Moreover, it was shown that STN7 kinase acts as a com-
mon redox sensor and/or signal transducer for both short-
and long-term responses in Arabidopsis thaliana (Pesaresi
et al. 2009), indicating that the two corresponding signal-
ling pathways diverge at, or immediately after downstream
of STN7. As suggested earlier (Allen and Pfannschmidt
2000), these two pathways may be subject to regulatory
coupling. It is noticeable that, in contrast to Arabidopsis,
no changes in photosystem stoichiometry could be detected
between the Chlamydomonas mutants stt7 and dum22
locked in state 1 and state 2, respectively, suggesting that
the Stt7 kinase is not involved in the readjustment of
photosystem stoichiometry (Cardol et al. 2009). Earlier
studies indicated that the PSI/PSII ratio changes in
39
Chlamydomonas cells grown under photoautotrophic con-
ditions in response to changes in the quality of irradiance
and that this chromatic regulation improves the quantum
efficiency of photosynthesis (Melis et al. 1996). The dif-
ferences observed between the roles of Stt7 and STN7 in
LTR may be due to structural differences between these
two proteins. While the N-terminal region and the catalytic
region are well conserved, the C-terminal domains differ
considerably and may have different functions.
Analysis of transcription patterns upon long-term accli-
mation using microarrays failed to reveal significant dif-
ferences between wild type and the stn7 mutant (Bonardi
et al. 2005; Tikkanen et al. 2006). However, the amount of
some thylakoid proteins was altered. In stn7, the level of
Lhcb1 decreased whereas an increases of Lhca1 and Lhca2
and of other chloroplast proteins were observed (Tikkanen
et al. 2006). In contrast, clear differences in the expression of
genes involved in photosynthesis or metabolism between
wild type and stn7 were observed during a time-course
experiment following a change to PSII light (Brautigam
et al. 2009). A large majority, 85% of the genes responding
to the light shift did no longer respond in stn7 indicating that
most of these genes are under STN7-mediated redox control
under these particular conditions. Moreover, under condi-
tions of fluctuating light intensity, the stn7 mutant displayed
retarded growth, and the expression of stress-responsive
genes was highly increased as compared to the wild type
(Tikkanen et al. 2006). It has, therefore, been proposed that
STN7 and state transitions are part of a buffering system
which dampens rapid oscillations of redox signals produced
in the photosynthetic apparatus by fluctuating environmen-
tal conditions. This allows the organisms to prevent over-
reduction of the electron acceptors of PSI which would
otherwise generate reactive oxygen species and induce the
expression of stress response genes (Tikkanen et al. 2006).
It also allows for the stabilization of metabolic fluxes and
coordinates photosynthesis and metabolism for adapting
plant growth to the environmental conditions (Brautigam
et al. 2009).
Interplay between kinases and phosphatases
Using high-accuracy mass spectrometry, the phospho-
proteome of Arabidopsis thaliana seedlings has been
characterized revealing four phosphorylation sites in the
C-terminal region of STN7 (Fig. 2) (Reiland et al. 2009).
Interestingly, this C-terminal region is not conserved
between Chlamydomonas Stt7 and Arabidopsis STN7
(Depège et al. 2003) suggesting that the C-terminal phos-
phorylation of STN7 controls responses that are specific to
higher plants. Furthermore, Reiland et al. (2009) proposed
that one phosphorylation site of STN7 may be a substrate
123
40
site for casein kinase 2 (CK2) which is known to phos-
phorylate proteins involved in transcription and post-
transcriptional regulation (Link 2003; Bollenbach et al.
2004). This raises the possibility that CK2 could be involved
in the phosphorylation processes in LHCII through the
STN7 kinase.
Stl1 is another protein kinase related to Stt7 in Chla-
mydomonas reinhardtii which is likely to be targeted to the
chloroplast because of the presence of an N-terminal
chloroplast presequence (Depège et al. 2003). However,
the function of this kinase is not yet known. Stl1 appears to
have an ortholog in Arabidopsis thaliana called STN8. The
availability of Arabidopsis T-DNA insertion lines with
disruptions in the STN7 and STN8 genes provided impor-
tant insights into the function of the corresponding pro-
teins. Both STN7 (Bellafiore et al. 2005) and STN8
(Bonardi et al. 2005; Vainonen et al. 2005) are activated in
thylakoid membranes by light. Loss of STN8 kinase leads
to a specific decrease in phosphorylation of the PSII core
proteins D1, D2 and CP43 (Bonardi et al. 2005; Vainonen
et al. 2005).
The existence of the conserved kinase couple STN7/
STN8 and Stt7/Stl1 in Arabidopsis and Chlamydomonas
raises the question of a possible functional interaction
between STN7/Stt7 and STN8/Stl1. Both proteins appear
to act synergistically based on the fact that the phenotype
of dephosphorylation of LHCII and of the PSII core pro-
teins in the double mutant stn7stn8 is more pronounced
than in the two single mutants together (Bonardi et al.
2005; Vainonen et al. 2005). Moreover, field tests revealed
that fitness as measured by seed production is significantly
decreased in the double mutant whereas it is decreased to a
smaller extent in stn7 and not significantly affected in stn8
(Frenkel et al. 2007). Also, the phosphorylation in state 2 of
Stl1 in Chlamydomonas is dependent on the presence of
the Stt7 kinase (Lemeille et al. 2010). In Arabidopsis, light
quality-dependent changes in core protein phosphorylation
do no longer occur in the stn7 mutant implying that STN7
affects activity or substrate accessibility of/for STN8 and
suggesting that as Stt7, STN7 acts upstream of STN8. The
synergistic functions of STN8 and SNT7 in Arabidopsis
were the basis for the proposal of the ‘‘sensor box’’ model
in which both kinases interact directly or indirectly to adapt
photosynthetic activity to changes in light quality and
quantity (Dietzel et al. 2008). The function of Stl1 in
Chlamydomonas may fulfil a similar function as STN8 in
Arabidopsis based on the sequence similarity. A major role
of this sensor box in C. reinhardtii most likely would be to
mediate the response to changes in metabolic state by
modulating the contributions of cyclic and linear electron
flow and thereby adjusting the ATP/NADPH ratio.
Recently, the chloroplast CSK kinase was identified and
proposed to act as a sensor kinase based on its homology to
123
Photosynth Res (2010) 106:33–46
bacterial two-component systems (Puthiyaveetil et al.
2008). It was proposed that autophosphorylation of CSK
occurs on Tyr because phosphorylation is resistant to acid
and alkali treatment. However, an extensive phosphopro-
teomic study did not reveal any chloroplast Tyr phos-
phorylation site (Reiland et al. 2009). CSK appears to be
conserved in most photosynthetic organisms although no
closely related kinase could be found in Chlamydomonas.
Chloroplast transcript accumulation was changed in CSK
knock-out lines indicating that CSK affects plastid gene
expression either directly or indirectly (Puthiyaveetil et al.
2008). The suggested redox regulatory coupling of CSK
between photosynthesis and chloroplast gene expression
requires further investigation to understand the functional
role of this kinase in detail.
Besides Stt7/STN7 and Stl1/STN8 which appear to be
linked through a phosphorylation cascade and the possible
connection between CK2 and STN7, it is not yet clear
whether there are links between these different kinases and
CSK which would involve other signalling chains in the
chloroplast (Fig. 3).
The reversible phosphorylation of the LHCII proteins
during state transitions implies the active participation of
protein phosphatases. Reversible phosphorylation of LHCII
proteins was observed with isolated thylakoids indicating
that at least a portion of the phosphatase is membrane
associated (Bennett 1980). It was further shown that the
Fig. 3 Thylakoid protein kinase network. The redox state of the
plastoquinone pool, which is influenced by light quality and quantity,
cellular ATP level and CO2 level, modulates the activity of the Stt7/
STN7 and Stl1/STN8 kinases required for the phosphorylation of
LHCII and PSII, respectively. It may also act on CSK. Activation of
the Stt7/STN7 kinase depends on its close interaction with the Cytb6f
complex. After it is phosphorylated, LHCII moves from PSII to PSI.
A phosphatase (Pase) which specifically dephosphorylates LHCII has
been identified recently (see text). Moreover, the redox state of Trx
also influences the activity of Stt7/STN7. The proposed action of CK2
on Stt7/STN7 and of CSK on chloroplast gene expression is still
tentative and will need further experimental support. Besides its role
in short-term acclimation, STN7 is also involved in the long-term
response (LTR). Protein kinases are highlighted in yellow
Photosynth Res (2010) 106:33–46
activities of thylakoid protein phosphatases are redox inde-
pendent and kinetically heterogeneous (Silverstein et al.
1993). Several chloroplast phosphatase activities were
identified both in thylakoid membranes and in the chloro-
plast stroma using as assays dephosphorylation of synthetic
phosphopeptide analogues of the LHCII N-terminal phos-
phorylation sites or authentic phosphopeptides cleaved from
thylakoid membrane proteins. In this way, a 29-kDa stromal
phosphatase was purified and suggested to play a role in the
dephosphorylation of LHCII (Hammer et al. 1997). How-
ever, its in vivo role remains unclear. It is not known whether
the LHCII phosphorylation state is solely regulated by the
LHCII kinase or whether phosphatases are also subjected to
regulation.
In contrast to soluble phosphatases which associate with
the thylakoid membrane and which appear to be involved in
LHCII dephosphorylation (Hammer et al. 1997; Kieleczawa
et al. 1992), a thylakoid membrane, PP2A protein phos-
phatase, was purified which was efficient in dephosphoryl-
ating PSII phosphoproteins and was regulated by the
immunophilin-like TLP40 protein localized in the thylakoid
lumen (Fulgosi et al. 1998; Vener et al. 2000). It was pro-
posed that signalling from TLP40 to the protein phosphatase
coordinates dephosphorylation and folding of thylakoid
membrane proteins.
During the screening of Arabidopsis lines with T-DNA
insertions in nuclear genes encoding putative chloroplast
phosphatases, two groups have recently identified a novel
phosphatase of A. thaliana, called TAP38/PPH1, which is
specifically required for the dephosphorylation of LHCII,
but not of the PSII core proteins under state 1 conditions
(Pribil et al. 2010; Shapiguzov et al. 2010). This phos-
phatase, which belongs to the family of monomeric PP2C
type phosphatases, is a chloroplast protein and is mainly
associated with the stromal lamellae of the thylakoid
membranes. Loss of TAP38/PPH1 leads to an increase in
the antenna size of photosystem I and impairs transition
from state 2 to state 1. Thus, phosphorylation and
dephosphorylation of LHCII appear to be specifically
mediated by the STN7 kinase–TAP38/PPH1 phosphatase
pair. These two proteins emerge as key players in the
adaptation of the photosynthetic apparatus to changes in
light quality and quantity.
Membrane remodelling during state transitions
A remarkable feature of state transitions is that they
involve considerable membrane remodelling. During a
transition from state 1 to state 2, spectroscopic measure-
ments indicate that 80% of the LHCII antenna of C. rein-
hardtii moves from PSII to PSI (Delosme et al. 1996). An
immunocytochemical study revealed marked changes in
41
the distribution of the cytochrome b6f complex and LHCII
between the appressed and unappressed thylakoid mem-
brane regions during state transitions (Vallon et al. 1991).
There was a significant enrichment of these two complexes
in the unappressed regions in state 2 compared to state 1.
Under similar conditions, this reorganization of LHCII and
the cytochrome b6f complex within the thylakoid mem-
branes was suppressed in the stt7 mutant which is locked in
state 1 (Fleischmann et al. 1999).
There is a marked difference in thylakoid membrane
organization between C. reinhardtii and land plants.
Whereas clear differences are observed in Arabidopsis
between grana regions and stromal lamellae, in C. rein-
hardtii, the grana regions generally consist of only a few
appressed membranes. A slightly higher extent of appres-
sed membranes was reported for C. reinhardtii cells in state
1 compared to state 2. Earlier studies concluded that the
unappressed regions carry a higher negative surface charge
than the appressed regions of thylakoid membranes (Barber
1982). It was shown that destacking of thylakoid mem-
branes can be induced in vitro through depletion of cations
presumably because of the appearance of repulsive nega-
tive charges. In contrast, readdition of cations promoted
restacking through screening of the negative surface char-
ges. The availability of mutants deficient in LHCII or PSII
core protein phosphorylation has made it possible to
investigate which surface charges are mainly responsible
for the folding of the thylakoid membranes. Electron
microscope analysis of the membranes from the stn7stn8
double mutant of Arabidopsis deficient in the light-induced
phosphorylation of LHCII and PSII core proteins revealed
a more compact thylakoid organization with increased
thylakoid membrane folding as compared to wild-type
plants (Fristedt et al. 2009). There was a significant
enhancement in the size of stacked thylakoid membranes in
stn7stn8 which was also manifested by a higher rate of
gravity-driven sedimentation of isolated thylakoids. This
phenotype was only seen in the absence of the STN8 kinase
but not in stn7 which lacks the STN7 kinase. Thus, PSII
core protein phosphorylation plays a critical role in the
folding of the thylakoid membranes. In contrast, the
phosphorylation status of LHCII does not appear to have a
significant effect on this process. The enhanced membrane
folding in stn7stn8 and stn8 hindered the lateral migration
of the PSII reaction centre protein D1 and of the processing
FtsH protease between the appressed and the non-appres-
sed membrane regions and slowed down D1 turnover in
plants exposed to high light (Fristedt et al. 2009). These
results suggest that the high level of PSII core protein
phosphorylation in plants is required for adjusting the
folding of the photosynthetic membranes for controlling
lateral mobility of membrane proteins and photosynthetic
activity.
123
42
Biochemical evidence for the displacement of LHCII
from PSII to PSI was provided through the isolation of a
PSI–LHCI–LHCII supercomplex from Arabidopsis plants
in state 2 (Zhang and Scheller 2004). Electron microscopy
revealed that this supercomplex consists of one PSI–LHCI
complex and one LHCII trimer (Kouril et al. 2005). The
docking site of LHCII on PSI is formed by the PsaH, PsaI
and PsaO subunits as revealed by cross-linking studies
which are in full agreement with the fact that mutants of
Arabidopsis lacking the PsaH, PsaL and PsaO subunits are
unable to perform state transitions (Lunde et al. 2000).
Recent experiments with Chlamydomonas also indicate a
clear association of LHCII to PSI under state 2 conditions.
A PSI–LHCI–LHCII supercomplex distinctly larger than
the PSI–LHCI complex was isolated which contains the
three LHCII proteins CP29, CP26 and Lhcbm5 (Takahashi
et al. 2006). The last-mentioned protein is a LHCII type II
protein which is present in similar amounts as CP29 and
CP26 but at a lower level than the other LHCII proteins.
Lhcbm5 may have a similar role as CP24 in land plants.
Together with CP29 and CP26, Lhcbm5 may act as a linker
between the PSII core dimer and the trimeric LHCII in
state 1. In state 2, these three proteins migrate to the PsaH
side of PSI where they provide a binding site for the LHCII
trimers on PSI. In this respect, it is interesting to note that
the crystal structure of the PSI–LHCI complex reveals that
four Lhca subunits form a belt on the opposite side of PsaH
(Ben-Shem et al. 2003). Thus, it appears that CP29, CP26
and Lhcbm5 shuttle between PSII and PSI during state
transitions and that their affinities for PSII and PSI may be
modulated by phosphorylation. Phosphorylated forms
of CP29 and Lhcbm5 were found associated with PSI
(Takahashi et al. 2006). Of particular interest is CP29 with
four sites phosphorylated in state 2 (Turkina et al. 2006).
The phosphorylation of three additional sites under high
light could induce its dissociation and that of LHCII from
PSI and allow for thermal energy dissipation within the
LHCII trimers. It thus appears that reversible phosphory-
lations at the interface between the PSII core and LHCII
play a critical role in state transitions.
In order to examine the dissociation of LHCII from the
PSII complex which occurs during a transition from state 1
to state 2, PSII-containing complexes were purified through
a His-tag inserted in CP47 by nickel affinity chromatog-
raphy (Iwai et al. 2008). This led to the identification of a
PSII core complex, a PSII–LHCII supercomplex, and a
multimer of PSII supercomplex called PSII megacomplex.
The fact that the megacomplex was predominant in state 1
while the core complex was found in state 2 indicated that
the LHCIIs dissociate from PSII on state transitions.
Moreover, phosphorylated LHCII type I was mainly found
in the supercomplex and less in the megacomplex whereas
phosphorylated CP26 and CP29 were only present in the
123
Photosynth Res (2010) 106:33–46
unbound form. Iwai et al. (2008) propose a model in which
PSII remodelling during transition from state 1 to state 2
proceeds in two steps; first a dissociation of the mega-
complex into supercomplexes mediated mainly by phos-
phorylation of LHCII type I and subsequent release of
LHCII from the supercomplex triggered by phosphoryla-
tion of the minor LHCIIs and the PSII core subunits. In this
view, the minor complexes CP29 and CP26 play a key role
as their phosphorylation induces the undocking of the
entire peripheral antenna during a transition to state 2. The
free LHCII would then reassociate with the PSI–LHCI
supercomplex. In order to further examine the role of CP29
and CP26 in state transitions in C. reinhardtii, RNA
interference experiments were performed (Tokutsu et al.
2009). The results indicate that, in the absence of CP29,
state transitions do no longer occur and although the
mobile LHCII is detached from PSII, it does not bind to the
PSI–LHCI complex. In contrast, the loss of CP26 did not
affect state transitions in this alga. This further confirms
that CP29 is essential for the docking of LHCII to PSI.
The traditional view of state transitions attributes an
important role to the movement of LHCII from PSII in the
grana to PSI in the stroma lamellae. A recent study has
compared the phosphorylation-dependent movement of
LHCII with the fluorescence changes occurring in both
photosystems upon subfractionation of the thylakoid
membranes by phase partitioning (Tikkanen et al. 2008).
Although several LHCII proteins moved to the lamellar
regions after LHCII phosphorylation, there was no increase
in PSI fluorescence associated with this fraction. Such an
increase in PSI fluorescence was exclusively found in the
grana margin fractions indicating that besides the mobile
LHCII, PSI–LHCI complexes appear to move from the
stromal lamellae to the grana margins where they associate
with LHCII and/or that a large fraction of PSI is located at
the margins as proposed by Albertsson (2001). In this view,
the dynamic nature of grana margins allows the capture of
excitation energy by PSI from PSII–LHCII in a process
which must be regulated in a tight and dynamic way
(Tikkanen et al. 2008).
Analysis of thylakoid membranes of Arabidopsis in state
1 by electron microscope tomography revealed that the
granum layers consist of repeating units that are physically
connected to each other and to the adjacent stroma lamellae
(Shimoni et al. 2005). According to this view, the grana
consist of bifurcations of stroma lamellae into neighbour-
ing sheets that run parallel to each other and that fuse with
each other through membrane bridges that stabilize the
membrane network. Reversible structural changes in thy-
lakoid membrane organization of this type have been
observed during state transitions by atomic force micros-
copy, scanning and transmission electron microscopy, and
confocal imaging (Chuartzman et al. 2008). Based on this
Photosynth Res (2010) 106:33–46
analysis, a model has been proposed in which reorganization
of the membranes is mediated through fusion and fission
events at the interface between the granal and stromal
lamellar domains of the thylakoid membranes. This mem-
brane remodelling appears to be initiated at the curved
margins of the appressed grana domains through breakage of
the lateral and vertical connections within this membrane
system. These localized rearrangements are proposed to
undergo macroscopic cooperative changes that then propa-
gate through the entire membrane network (Chuartzman
et al. 2008). A challenging task will be to identify and
characterize the intrachloroplast machinery which partici-
pates in these fission and fusion events.
Physiological significance of state transitions
It is apparent that state transitions in C. reinhardtii are
associated with the movement of a large part of the LHCII
antenna. In this organism, state transitions are influenced to
a large extent by the cellular ATP level. Lack of ATP
results in a transition to state 2. Moreover, a transition from
state 1 to state 2 in this alga acts as a switch from linear to
cyclic electron flow and, therefore, influences the ATP/
NADPH ratio (Finazzi et al. 1999, 2002). It is very likely
that the major role of state transitions in this alga is to
adjust the cellular ATP levels in response to changes in
cellular demand rather than the balancing of the light
excitation energy between the two photosystems. It would
be difficult to explain the migration of 80% of the LHCII
antenna from PSII to PSI uniquely for reasons of antenna
size readjustment.
It is surprising that no obvious growth phenotype could
be detected in the stt7 mutant of C. reinhardtii even under
changing light conditions (Fleischmann et al. 1999).
However, the situation changes drastically under condi-
tions leading to low ATP levels. It was noticed that
depletion of ATP through inhibition of mitochondrial res-
piration or in mitochondrial mutants deficient in respira-
tion, in the dark, stimulates glycolysis through the Pasteur
effect (Bulté et al. 1990; Cardol et al. 2003). This, in turn,
leads to the production of reductants which reduce the
plastoquinone pool and induce state 2. Transfer of these
cells to the light restores ATP levels and state 1 partially.
These findings suggested a possible link between state
transitions and mitochondrial respiration. In order to fur-
ther explore the interplay between state transitions and
mitochondrial respiration, the growth properties of the stt7
mutant were examined in the presence of myxothiazol, an
inhibitor of respiration (Cardol et al. 2003). The growth of
this mutant on minimal medium was drastically affected.
Next, the double mutant stt7 dum22 deficient both in state
43
transitions and mitochondrial respiration was examined. Its
growth rate under photoautotrophic conditions was shown
to be severely affected at low light but less so under high
light, indicating that the growth defect is not due to
increased photosensitivity. These results indicate that
Chlamydomonas cells display some energetic flexibility
and are able to compensate for a lack of mitochondrial
respiration and ATP synthesis through transition to state 2
and a concomitant switch to cyclic electron flow which
enhances ATP production for sustained CO2 assimilation.
Indeed, an ATP to NADPH ratio of 1:5 is required to drive
the Calvin–Benson cycle which cannot be achieved solely
by linear electron flow. In the absence of state transitions,
mitochondrial respiration can, at least, partially compen-
sate. However, under conditions of impaired mitochondrial
respiration, state transitions are essential for ensuring
photoautotrophic growth under limiting light. This is
because, in mitochondrial respiration mutants, increased
reducing power is fed into the chloroplast plastoquinone
pool together with the electrons originating from PSII. This
increase must be matched by an increase in the PSI
absorption capacity through a transition to state 2, thereby
allowing for an efficient light utilization by both photo-
systems. Thus, there is a tight interplay between cyclic
electron flow and respiratory capacity which is mediated, in
part, through the process of state transitions.
The importance of state transitions is apparent not only
in Chlamydomonas but also in Arabidopsis. The analysis of
double mutants of Arabidopsis combining the stn7 muta-
tion with mutations affecting linear electron flow and
leading to a more reduced state of the plastoquinone pool
showed that the growth rate and the effective quantum
yield were significantly decreased compared to the single
mutants (Pesaresi et al. 2009). These results indicate that
state transitions become critical when linear electron
transport is impaired. Comparison of the stn7 mutant with
wild-type Arabidopsis lines in field tests revealed that the
loss of state transitions leads to a significant decrease in
fitness (Frenkel et al. 2007). Moreover, controlled light
shift experiments with stn7 and wild-type plants also
showed marked differences in seed production (Wagner
et al. 2008). Thus, when the plants were shifted every
2–3 days between PSI- and PSII-light, a period which
allows for LTR, stn7 plants produced 50% less seeds than
wild-type plants. When the light shifts occurred every
20 min which correspond to the time range of state tran-
sitions, a further decrease in seed production was observed
for stn7. It, thus, appears that state transitions and LTR
provide a metabolic flexibility which allows photosynthetic
organisms to acclimate both under long- and short-term
light quality shifts corresponding to a wide range of light
fluctuations which occur in a natural environment.
123
44
Acknowledgements We thank Michel Goldschmidt-Clermont for
his helpful comments. Research in the authors’ laboratory was sup-
ported by a grant from the Swiss National Foundation (3100AO-
117712) to J.D.R.
References
Adam Z, Clarke AK (2002) Cutting edge of chloroplast proteolysis.
Trends Plant Sci 7:451–456
Albertsson PA (2001) A quantitative model of the domain structure of
the photosynthetic membrane. Trends Plant Sci 6:349–354
Allen JF (1992) Protein phosphorylation in regulation of photosyn-
thesis. Biochim Biophys Acta 1098:275–335
Allen JF (2003) Cyclic, pseudocyclic and noncyclic photophosphor-
ylation: new links in the chain. Trends Plant Sci 8:15–19
Allen JF, Pfannschmidt T (2000) Balancing the two photosystems:
photosynthetic electron transfer governs transcription of reaction
centre genes in chloroplasts. Philos Trans R Soc Lond B Biol Sci
355:1351–1359
Allen JF, Bennett J, Steinback KE, Arntzen CJ (1981) Chloroplast
protein phosphorylation couples plastoquinone redox state to
distribution of excitation energy between photosystems. Nature
291:25–29
Andersson B, Andersson J (1980) Lateral heterogeneity in the
distribution of chlorophyll–protein complexes of the thylakoid
membranes of spinach chloroplasts. Biochem Biophys Acta
593:427–440
Barber J (1982) The control of membrane organization by electro-
static forces. Biosci Rep 2:1–13
Bellafiore S, Barneche F, Peltier G, Rochaix JD (2005) State
transitions and light adaptation require chloroplast thylakoid
protein kinase STN7. Nature 433:892–895
Bennett J (1977) Phosphorylation of chloroplast membrane proteins.
Nature 269:344–346
Bennett J (1979) Chloroplast phosphoproteins. Phosphorylation of
polypeptides of the light-harvesting chlorophyll protein com-
plex. Eur J Biochem 99:133–137
Bennett J (1980) Chloroplast phosphoproteins. Evidence for a
thylakoid-bound phosphoprotein phosphatase. Eur J Biochem
104:85–89
Ben-Shem A, Frolow F, Nelson N (2003) Crystal structure of plant
photosystem I. Nature 426:630–635
Bollenbach TJ, Schuster G, Stern DB (2004) Cooperation of endo-
and exoribonucleases in chloroplast mRNA turnover. Prog
Nucleic Acid Res Mol Biol 78:305–337
Bonardi V, Pesaresi P, Becker T, Schleiff E, Wagner R, Pfannschmidt
T, Jahns P, Leister D (2005) Photosystem II core phosphoryla-
tion and photosynthetic acclimation require two different protein
kinases. Nature 437:1179–1182
Bonaventura C, Myers J (1969) Fluorescence and oxygen evolution
from Chlorella pyrenoidosa. Biochim Biophys Acta 189:366–383
Brautigam K, Dietzel L, Kleine T, Stroher E, Wormuth D, Dietz KJ,
Radke D, Wirtz M, Hell R, Dormann P, Nunes-Nesi A, Schauer N,
Fernie AR, Oliver SN, Geigenberger P, Leister D, Pfannschmidt T
(2009) Dynamic plastid redox signals integrate gene expression
and metabolism to induce distinct metabolic states in photosyn-
thetic acclimation in Arabidopsis. Plant Cell 21:2715–2732
Breyton C (2000) Conformational changes in the cytochrome b6f
complex induced by inhibitor binding. J Biol Chem 275:13195–
13201
Bulté L, Gans P, Rebeille F, Wollman FA (1990) ATP control on state
transitions in Chlamydomonas. Biochim Biophys Acta 1020:
72–80
123
Photosynth Res (2010) 106:33–46
Cardol P, Gloire G, Havaux M, Remacle C, Matagne R, Franck F
(2003) Photosynthesis and state transitions in mitochondrial
mutants of Chlamydomonas reinhardtii affected in respiration.
Plant Physiol 133:2010–2021
Cardol P, Alrich J, Girard-Bascou J, Franck F, Wollman FA, Finazzi
G (2009) Impaired respiration discloses the physiological
significance of state transitions in Chlamydomonas. Proc Natl
Acad Sci USA 106:15979–15984
Chuartzman SG, Nevo R, Shimoni E, Charuvi D, Kiss V, Ohad I,
Brumfeld V, Reich Z (2008) Thylakoid membrane remodeling
during state transitions in Arabidopsis. Plant Cell 20:1029–1039
Cleland RE, Bendall DS (1992) Photosystem I cyclic electron
transport: measurement of ferredoxin-plastoquinone reductase
activity. Photosynth Res 34:409–418
Coughlan SJ, Hind G (1986) Purification and characterization of a
membrane-bound protein kinase from spinach thylakoids. J Biol
Chem 261:11378–11385
Coughlan S, Hind G (1987) Phosphorylation of thylakoid proteins by
a purified kinase. J Biol Chem 262:8402–8408
DalCorso G, Pesaresi P, Masiero S, Aseeva E, Schunemann D,
Finazzi G, Joliot P, Barbato R, Leister D (2008) A complex
containing PGRL1 and PGR5 is involved in the switch between
linear and cyclic electron flow in Arabidopsis. Cell 132:273–285
de Lacroix de Lavalette A, Finazzi G, Zito F (2008) b6f-Associated
chlorophyll: structural and dynamic contribution to the different
cytochrome functions. Biochemistry 47:5259–5265
Delosme R, Olive J, Wollman FA (1996) Changes in light energy
distribution upon state transitions: an in vivo photoacoustic study
of the wild type and photosynthesis mutants from Chlamydo-
monas reinhardtii. Biochim Biophys Acta 1273:150–158
Depège N, Bellafiore S, Rochaix JD (2003) Role of chloroplast
protein kinase Stt7 in LHCII phosphorylation and state transition
in Chlamydomonas. Science 299:1572–1575
Dietzel L, Bräutigam K, Pfannschmidt T (2008) Photosynthetic
acclimation: state transitions and adjustment of photosystem
stoichiometry–functional relationships between short-term
and long-term light quality acclimation in plants. FEBS J 275:
1080–1088
Finazzi G, Furia A, Barbagallo RP, Forti G (1999) State transitions,
cyclic and linear electron transport and photophosphorylation
in Chlamydomonas reinhardtii. Biochim Biophys Acta 1413:
117–129
Finazzi G, Zito F, Barbagallo RP, Wollman FA (2001) Contrasted
effects of inhibitors of cytochrome b6f complex on state
transitions in Chlamydomonas reinhardtii: the role of Qo site
occupancy in LHCII kinase activation. J Biol Chem 276:9770–
97743
Finazzi G, Rappaport F, Furia A, Fleischmann M, Rochaix JD, Zito F,
Forti G (2002) Involvement of state transitions in the switch
between linear and cyclic electron flow in Chlamydomonas
reinhardtii. EMBO Rep 3:280–285
Fleischmann MM, Ravanel S, Delosme R, Olive J, Zito F, Wollman
FA, Rochaix JD (1999) Isolation and characterization of
photoautotrophic mutants of Chlamydomonas reinhardtii defi-
cient in state transition. J Biol Chem 274:30987–30994
Frenkel M, Bellafiore S, Rochaix JD, Jansson S (2007) Hierarchy
amongst photosynthetic acclimation responses for plant fitness.
Physiol Plantarum 129:455–459
Fristedt R, Willig A, Granath P, Crèvecoeur M, Rochaix JD, Vener A
(2009) Phosphorylation of photosystem II controls functional
macroscopic folding of plant photosynthetic membranes. Plant
Cell 21:3950–3964
Fulgosi H, Vener AV, Altschmied L, Herrmann RG, Andersson B
(1998) A novel multi-functional chloroplast protein: identifica-
tion of a 40 kDa immunophilin-like protein located in the
thylakoid lumen. EMBO J 17:1577–1587
Photosynth Res (2010) 106:33–46
Gal A, Hauska G, Herrmann RG, Ohad I (1990) Interaction between
light-harvesting chlorophyll-a/b protein (LHCII)kinase and
cytochrome b6f complex. In vitro control of kinase activity.
J Biol Chem 265:17749–19742
Hamel P, Olive J, Pierre Y, Wollman FA, de Vitry C (2000) A new
subunit of cytochrome b6f complex undergoes reversible
phosphorylation upon state transition. J Biol Chem 275:17072–
17079
Hammer MF, Markwell J, Sarath G (1997) Purification of a protein
phosphatase from chloroplast stroma capable of dephosphoryl-
ating the light-harvesting complex-II. Plant Physiol 113:227–233
Helm M, Luck C, Prestele J, Hierl G, Huesgen PF, Frohlich T, Arnold
GJ, Adamska I, Gorg A, Lottspeich F, Gietl C (2007) Dual
specificities of the glyoxysomal/peroxisomal processing protease
Deg15 in higher plants. Proc Natl Acad Sci USA 104:11501–
11506
Holt NE, Fleming GR, Niyogi KK (2004) Toward an understanding
of the mechanism of nonphotochemical quenching in green
plants. Biochemistry 43:8281–8289
Horton P, Black MT (1980) Activation of adenosine 50 triphosphate-
induced quenching of chlorophyll fluorescence by reduced
plastoquinone. FEBS Lett 119:141–145
Hou CH, Rintamäki E, Aro EM (2003) Ascorbate-mediated LHCII
protein phosphorylation—LHCII kinase regulation in light and
in darkness. Biochemistry 42:5828–5836
Huesgen PF, Schuhmann H, Adamska I (2009) Deg/HtrA proteases as
components of a network for photosystem II quality control in
chloroplasts and cyanobacteria. Res Microbiol 160:726–732
Iwai M, Takahashi Y, Minagawa J (2008) Molecular remodeling of
photosystem II during state transitions in Chlamydomonas
reinhardtii. Plant Cell 20:2177–2189
Joliot P, Joliot A (2006) Cyclic electron flow in C3 plants. Biochim
Biophys Acta 1757:362–368
Kieleczawa J, Coughlan SJ, Hind G (1992) Isolation and character-
ization of an alkaline phosphatase from pea thylakoids. Plant
Physiol 99:1029–1036
Kouril R, Zygadlo A, Arteni AA, DeWit CD, Delkker JP, Jensen PE,
Scheller HV, Boekema EJ (2005) Structural characterization of a
complex of photosystem I and light-harvesting complex II of
Arabidopsis thaliana. Biochemistry 44:10935–10940
Kruse O, Nixon PJ, Schmid GH, Mullineaux CW (1999) Isolation of
state transition mutants of Chlamydomonas reinhardtii by
fluorescence video imaging. Photosynth Res 61:43–51
Lemeille S, Willig A, Depège-Fargeix N, Delessert C, Bassi R,
Rochaix JD (2009) Analysis of the chloroplast protein kinase
Stt7 during state transitions. PLoS Biol 7:e45
Lemeille S, Turkina M, Vener AV, Rochaix JD (2010) Stt7-
dependent phosphorylation during state transitions in the green
alga Chlamydomonas reinhardtii. Mol Cell Proteomics. doi:
10.1074/mcp.M000020-MCP201
Lennartz K, Plucken H, Seidler A, Westhoff P, Bechtold N, Meierhoff
K (2001) HCF164 encodes a thioredoxin-like protein involved in
the biogenesis of the cytochrome b(6)f complex in Arabidopsis.
Plant Cell 13:2539–2551
Link G (2003) Redox regulation of chloroplast transcription. Antioxid
Redox Signal 5:79–87
Lunde C, Jensen PE, Haldrup A, Knoetzel J, Scheller HV (2000) The
PSI-H subunit of photosystem I is essential for state transitions in
plant photosynthesis. Nature 408:613–615
Melis A, Murakami A, Nemson JA, Aizawa K, Ohki K, Fujita Y
(1996) Chromatic regulation in Chlamydomonas reinhardtii
alters photosystem stoichiometry and improves the quantum
efficiency of photosynthesis. Photosynth Res 47:253–265
Moss DA, Bendall DS (1984) Cyclic electron transport in chloro-
plasts: the Q-cycle and the site of action of anthocyanin.
Biochim Biophys Acta 767:389–395
45
Motohashi K, Hisabori T (2006) HCF164 receives reducing equiv-
alents from stromal thioredoxin across the thylakoid membrane
and mediates reduction of target proteins in the thylakoid lumen.
J Biol Chem 281:35039–35047
Munekage Y, Hojo M, Meurer J, Endo T, Tasaka M, Shikanai T
(2002) PGR5 is involved in cyclic electron flow around
photosystem I and is essential for photoprotection in Arabidop-
sis. Cell 110:361–371
Murata N (1969) Control of excitation transfer in photosynthesis. I.
Light-induced change of chlorophyll a fluorescence in Porphyri-
dium cruentum. Biochim Biophys Acta 172:242–251
Page ML, Hamel PP, Gabilly ST, Zegzouti H, Perea JV, Alonso JM,
Ecker JR, Theg SM, Christensen SK, Merchant S (2004) A
homolog of prokaryotic thiol disulfide transporter CcdA is
required for the assembly of the cytochrome b6f complex in
Arabidopsis chloroplasts. J Biol Chem 279:32474–32482
Pesaresi P, Hertle A, Pribil M, Kleine T, Wagner R, Strissel H,
Ihnatowicz A, Bonardi V, Scharfenberg M, Schneider A,
Pfannschmidt T, Leister D (2009) Arabidopsis STN7 kinase
provides a link between short- and long-term photosynthetic
acclimation. Plant Cell 21:2402–2423
Pfannschmidt T (2003) Chloroplast redox signals: how photosynthesis
controls its own genes. Trends Plant Sci 8:33–41
Pribil M, Pesaresi P, Hertle A, Barbato R, Leister D (2010) Role of
plastid protein phosphatase TAP38 in LHCII dephosphorylation
an thylakoid electron flow. PLoS Biol 8:e 1000288
Puthiyaveetil S, Kavanagh TA, Cain P, Sullivan JA, Newell CA, Gray
JC, Robinson C, van der Giezen M, Rogers MB, Allen JF (2008)
The ancestral symbiont sensor kinase CSK links photosynthesis
with gene expression in chloroplasts. Proc Natl Acad Sci USA
105:10061–10066
Reiland S, Messerli G, Baerenfaller K, Gerrits B, Endler A,
Grossmann J, Gruissem W, Baginsky S (2009) Large-scale
Arabidopsis phosphoproteome profiling reveals novel chloro-
plast kinase substrates and phosphorylation networks. Plant
Physiol 150:889–903
Rintamaki E, Salonen M, Suoranta UM, Carlberg I, Andersson B, Aro
EM (1997) Phosphorylation of light-harvesting complex II and
photosystem II core proteins shows different irradiance-depen-
dent regulation in vivo. Application of phosphothreonine anti-
bodies to analysis of thylakoid phosphoproteins. J Biol Chem
272:30476–30482
Rochaix JD (2007) Role of thylakoid protein kinases in photosyn-
thetic acclimation. FEBS Lett 581:2768–2775
Shapiguzov A, Ingelsson B, Samol I, Andres C, Kessler F, Rochaix
JD, Vener AV, Goldschmidt-Clermont M (2010) The PPH1
phophatase is specifically involved in LHCII dephosphorylation
and state transitions in Arabidopsis. Proc Natl Acad Sci USA.
doi:10.1073/pnas.0913810107
Shikanai T (2007) Cyclic electron transport around photosystem I:
genetic approaches. Annu Rev Plant Biol 58:199–217
Shimoni E, Rav-Hon O, Ohad I, Brumfeld V, Reich Z (2005) Three-
dimensional organization of higher-plant chloroplast thylakoid
membranes revealed by electron tomography. Plant Cell
17:2580–2586
Silverstein T, Cheng L, Allen JF (1993) Chloroplast thylakoid protein
phosphatase reactions are redox-independent and kinetically
heterogeneous. FEBS Lett 334:101–105
Snyders S, Kohorn BD (1999) TAKs, thylakoid membrane protein
kinases associated with energy transduction. J Biol Chem
274:9137–9140
Snyders S, Kohorn BD (2001) Disruption of thylakoid-associated
kinase 1 leads to alteration of light harvesting in Arabidopsis.
J Biol Chem 276:32169–32176
Sokolenko A, Fulgosi H, Gal A, Altschmied L, Ohad I, Herrmann RG
(1995) The 64 kDa polypeptide of spinach may not be the LHCII
123
46
kinase, but a lumen-located polyphenol oxidase. FEBS Lett
371:176–180
Stroebel D, Choquet Y, Popot JL, Picot D (2003) An atypical haem in
the cytochrome b(6)f complex. Nature 426:413–418
Takahashi H, Iwai M, Takahashi Y, Minagawa J (2006) Identification
of the mobile light-harvesting complex II polypeptides for state
transitions in Chlamydomonas reinhardtii. Proc Natl Acad Sci
USA 103:477–482
Tikkanen M, Piippo M, Suorsa M, Sirpio S, Mulo P, Vainonen J,
Vener AV, Allahverdiyeva Y, Aro EM (2006) State transitions
revisited—a buffering system for dynamic low light acclimation
of Arabidopsis. Plant Mol Biol 62:779–79371
Tikkanen M, Nurmi M, Suorsa M, Danielsson R, Mamedov F, Styring
S, Aro EM (2008) Phosphorylation-dependent regulation of
excitation energy distribution between the two photosystems in
higher plants. Biochim Biophys Acta 1777:425–432
Tokutsu R, Iwai M, Minagawa J (2009) CP29, a monomeric light-
harvesting complex II protein, is essential for state transitions in
Chlamydomonas reinhardtii. J Biol Chem 284:7777–7782
Turkina MV, Kargul J, Blanco-Rivero A, Villarejo A, Barber J, Vener
AV (2006) Environmentally modulated phosphoproteome of
photosynthetic membranes in the green alga Chlamydomonas
reinhardtii. Mol Cell Proteomics 5:1412–1425 [Epub 2006 May 2]
Vainonen JP, Hansson M, Vener AV (2005) STN8 protein kinase in
Arabidopsis thaliana is specific in phosphorylation of photosys-
tem II core proteins. J Biol Chem 280:33679–33686
Vallon O, Bulte L, Dainese P, Olive J, Bassi R, Wollman FA (1991)
Lateral redistribution of cytochrome b6/f complexes along
thylakoid membranes upon state transitions. Proc Natl Acad
Sci USA 88:8262–8266
Vener AV, van Kan PJ, Rich PR, Ohad II, Andersson B (1997)
Plastoquinol at the quinol oxidation site of reduced cytochrome
bf mediates signal transduction between light and protein
phosphorylation: thylakoid protein kinase deactivation by a
single-turnover flash. Proc Natl Acad Sci USA 94:1585–1590
123
Photosynth Res (2010) 106:33–46
Vener AV, Rokka A, Fulgosi H, Andersson B, Herrmann RG (1999) A
cyclophilin-regulated PP2A-like protein phosphatase in thylakoid
membranes of plant chloroplasts. Biochemistry 38:14955–14965
[erratum appears in Biochemistry (2000) 39:2130]
Wagner R, Dietzel L, Bräutigam K, Fischer W, Pfannschmidt T
(2008) The long-term response to fluctuating light quality is an
important and distinct light acclimation mechanism that supports
survival of Arabidopsis thaliana under low light conditions.
Planta 228:573–587
Wollman FA (2001) State transitions reveal the dynamics and
flexibility of the photosynthetic apparatus. EMBO J 20:3623–
3630
Wollman FA, Delepelaire P (1984) Correlations between fluorescence
and phosphorylation changes in thylakoid membranes of Chla-
mydomonas reinhardtii in vivo: a kinetic analysis. J Cell Biol
98:1–7
Wollman FA, Lemaire C (1988) Studies on kinase-controlled state
transitions in photosystem II and b6f mutants from Chlamydo-
monas reinhardtii which lack quinone-binding proteins. Biochim
Biophys Acta 933:85–94
Zhang S, Scheller HV (2004) Light-harvesting complex II binds to
several small subunits of photosystem I. J Biol Chem 279:3180–
3187
Zhang Z, Huang L, Shulmeister VM, Chi YI, Kim KK, Hung LW,
Crofts AR, Berry EA, Kim SH (1998) Electron transfer by
domain movement in cytochrome bc1. Nature 392:677–684
Zito F, Finazzi G, Delosme R, Nitschke W, Picot D, Wollman FA
(1999) The Qo site of cytochrome b6f complexes controls the
activation of the LHCII kinase. EMBO J 18:2961–2966
Zito F, Vinh J, Popot JL, Finazzi G (2002) Chimeric fusions of
subunit IV and PetL in the b6f complex of Chlamydomonas
reinhardtii: structural implications and consequences on state
transitions. J Biol Chem 277:12446–12455